2, 4-diamino-6-non-oxo-carbonylic triazines



United States Patent 3,271,393 2,4-DIAMHN@-6-NON-0X0-CARBNYMC TRIAZINESTalteo Uetla, 1066 lzurni, Komae-cho, Kitatama-gun,

Tokyo, Japan; Shigeshi Toyoshima, 35S Funahashi-cho, Sedagaya-lru,Tokyo, Japan; and Mitsuru Furukawa, 897 Yukigaya-cho, (Eta-kn, Tokyo,Japan No Drawing. Filed Mar. 28, 1963, er. No. 268,551 Ciaims priority,application Japan, July 3, 1962, 37 27,619 4 Qlaims. (Cl. Zea-247.2

wherein R and R are members selected from the group consisting ofhydrogen, alkyl, alkenyl, aryl, aralkyl and cycloalkyl, and wherein thegroup taken together may be a heterocyclic group containing from 4 to 5carbon atoms; and Y is a member selected from the group consisting ofmorpholino, piperidino, pyr- 4 rolidino,

in which R and R are members selected from the group consisting ofhydrogen, alkyl, alkenyl, aralkyl and aryl. The group taken together maybe, for example, morpholino piperidino ice In a preferred embodiment ofthis invention, Y in Formula I is and pyrrolidino NIIG-NH:

l IH

and the group taken together is morpholino, i.e.,

The compounds of the present invention (except those wherein Y is OH)may be prepared by the reaction of an a-substituted biguanide having theformula R NH NI'I II II N-CNH-CNH wherein R and R are the same asdefined above, with a di(lower)a1kyl oxalate, such as diethyl oxalate.The reaction may be conducted in an inert reaction media such asmethanol. The reaction may be hastened by heating. This reaction resultsin the formation of a compound having the formula This product is thenreacted with ammonia or an amine selected from the group consisting ofNH2( 3-NHz NH and R3 NH in which R and R are the same as defined above,preferably in an inert reaction medium such as ethanol. The reaction ispreferably conducted at elevated temperatures. The product, representedby Formula I above, may be recovered by filtration. Alternatively, thesecompounds may be prepared in one step by reacting in the absence of anyadditional solvent an Ot-SUbStitUted biguanide having the Formula 11above, a di(lower)alkyl oxalate and ammonia or an amine selected fromthe group consisting of NII fiNII and in which R and R are the same asdefined above.

The compounds represented by Formula II are known in the art. Forexample, when R and R are both hydrogen, the compound is biguanide whichis commercially available. When the group taken together is morpholino,i.e.,

volves the reaction of a compound having the formula (I fl) (J-X /C\ t\NO o-mn R2 N in which R and R are the same as defined above and X is amember selected from the group consisting of chloro, bromo, fluoro,iodo, and OR (R being alkyl), with ammonia or an amine selected from thegroup consisting of and in which R and R are the same as defined above.Compounds of Formula IV in which X is OR may be obtained by the reactionof a compound of Formula III with an alcohol; or by the reaction of acompound of Formula II, a di(lower)alkyl oxalate and an alcohol.

Compounds represented by Formula I wherein Y is hydroxyl may besynthesized by the hydrolysis of a compound of Formula IV in which X isOR or by the hydrolysis of a compound of Formula III, or by reacting acompound of Formula II with an oxalic acid monoester. The product fromany of these reactions may be further reacted with a halogenationreagent to obtain a compound of Formula IV in which X is halogen.

The compounds of this invention possess useful antiviral activity,particularly vir ucidal activity. Some of the compounds, particularly2-amino-4-morpholino 6-(N- amidinocarbamoyl)-s-triazine are useful ininhibiting the multiplication of and inactivating of such strains ofvirus as the Mahoney and MEF strain of poliomyelitis virus and in thesuspension of the production of the toxin-like substance (the OPE.factor) produced by poliomyelitis virus. The compounds of the presentinvention also are effective in inhibiting the multiplication of themeasles virus (Edmonstan strain). They have also been found to have asignificant chemotherapeutic and chemoprophylactic effect on influenzavirus and to be effective against echo and coxsackie viruses. Thecompounds of this invention are also useful in the study of viruses,particularly as a tool to facilitate study of their basic biologicalcharacteristics, including their intracellular multiplication.

The compounds of the present invention may be associated with asignificant amount of a pharmaceutically acceptable carrier which may beeither a solid material or a liquid. The compositions may take the formof tablets, effervescent tablets, powder, granules, capsules (both hardand soft shell capsules), of suspensions in edible oils, or other dosageforms which are particularly useful for oral ingestion. Liquid diluentsare employed in sterile conditions for parenteral use, that is byintramuscular, intravenous and intraperitoneal injection. Such a mediummay be a sterile solvent or suspending agent such as water or aninjectable oil. The compositions may take the form of active materialadmixed with solid diluents and/or tabletting adjuvants such ascornstarch, lactose,

talc, stearic acid, magnesium stearate, gums or the like. Any of theencapsulating or tabletting materials used in pharmaceutical practicemay be employed where there is no .incompartibility with the compounds.The materials may be tabletted with or without adjuvants. Alternatively,the compounds may be placed in the usual capsule or resorbable materialsuch as the usual gelatin capsule and administered in that form. In yetanother embodiment, the compounds may be put up as a powder and soemployed, as by nasal inhalation, or the compounds may be prepared inthe form of a palatable suspension in which the compounds are notsoluble. Suspensions may be given orally as made or may be encapsulated.Ointments and lotions are useful topically; use for topical therapy ismade of nose drops, troches and suppositories. The compounds of thepresent invention are particularly useful when given by the oral,intramuscular, or intravenous routes.

The following examples will illustrate the preparation of the compoundsof the present invention.

EXAMPLE 1 A solution of sodium methoxide (0.1 mole) in absolute methanolis added, with heating and stirring, to a suspension of 0.1 mole ofpowdered 4-morpholinecarboximidoylguanidine hydrochloride in absolutemethanol. NaCl which is formed as a precipitate is removed byfiltration. Diethyl oxalate (0.1 mole) is then added and the solution isrefluxed for 30 minutes on a steam bath. The reaction which occurs maybe represented by the following equation:

The reaction mixture is allowed to stand overnight and the precipitatedproduct is collected by filtration. The prod uct is found to have amelting point of 268 C., a yield of 79%. Elemental analysis.Calculatedfor C H O N N, 31.10%. Found: 31.02%.

A suspension of 0.01 mole of the product obtained above is refluxed incc. of absolute ethanol until all of the material has been dissolved. Tothe solution there is added 0.01 mole of guanidine and the mixture isrefluxed for 5 hours. A white precipitate is formed during therefluxing. The reaction which occurs may be represented by the followingequation:

The reaction mixture is concentrated, the precipitate recovered andrecrystallized from water. The product, 2- amino-4-morpholino-6(N-amidinocarbamoyl)s-triazine, consists of prisms having a meltingpoint of 271 C. Elemental analysis.-Calculated for C H O N N, 42.08%.Found: N, 41.50%.

The inhibitory effect of 2-amino-4-morpholino-6-(N-amidinocarbamoyl)s-triazine on the multiplication of polio virus (type1Mahoney and type 2-MEF and measles (Edmonstan strain) is determinedusing the standard tissue culture method. KB cells were used for polioand Hep. No. 2 cells for measles. Table I sets forth the tissue cultureinhibitory dose (TCID) m1, for the compound tested as well as for thecontrol.

TABLE I TCIDao/o-r ml. Viral Material Control Tteated Polio:

Type 1Mahoney c 10c Type 2-MEF 100 106 Measles:

Edmonstan 10c 10c In another test procedure, the minimum inhibitoryconcentration of 2-amino-4-morpholino-6-(N-amidinocarbamoyl)-s-triazineon measles virus in Hep. No. 2 cells is determined. The results are setforth in Table 11.

TABLE II Concentration of compound (in moles): Results 10* 4/4 4 10 4/42 1O- 0/4 None 0/4 (In the results column, the denominator denotes thetotal number of assay tubes used in the experiment and the nu meratordenotes the number of tubes in which no cytopathogenic efiect wasobserved.)

The minimum inhibitory concentration of 2-amino-4- morpholino--(Namidinocarbamoyl)-s-triazine on the 6 EXAMPLE 3 Preparation of 1,1-(2,2-0xydie1hyl) -3-(4,5-di0x0-2- imidazolidinylidene) -gnania'ineEXAMPLE 4 Preparation of l-(p-merlloxyphenyl)-3-(4,5-di0xo-2-imidazolidinylidene)-guanidine NH NIT-0:0

To a solution of 2.07 g. of l-(p-methoxyphenyl)-biguanide in 5 cc. ofdehydrated methanol is added 1.46 g. of diethyl oxalate. After allowingto stand for 2 hours, the yellow precipitate immediately deposited fromthe re action mixture is collected by filtration; yield 2.40 g. (92%),M.P. 208 C. (with decomposition). Analysis.- Calcd. for C H O N N,26.81. Found: N, 27.05.

Other examples of intermediates of the type represented by Formula IIIwhich are prepared by the general method described above are set forthin Table III.

TABLE III R1 Analysis N Percent Melting Yield Molecular Example Group Nof formula III Point (Percent) Formula (0 C.) Cale. Exper. R2

5 NH? 300 C4H O2N5 45.15 45.

6 iS0-C3H7NH- 218 77 C7H1 O2N5 35. 52 35. 90

7 -NH- 249 83 C10Hl502N5- 29. 52 29.80

8 CH3 -NH- 214 95 C11HuO2N 28. 56 28.80

Mahoney strain of virus (10*) is found to be 4 10 mole.

The following examples are illustrative of the preparation of variousintermediates of the type represented for Formula 111, above:

EXAMPLE 2 Preparation 0 1,Z-dimellzyl-3-(4,5-di0x0-2-imidazolidinylidene)-gnanidine CH3 TH /NIIC=O NCN=C C63 \NI-I-C=O Sodiumethoxide prepared from 0.23 g. of metallic sodium and a small amount ofdehydrated ethanol is added, with warming, to a suspension of 1.65 g. of1,1-dimethylbiguanide hydrochloride in 5 cc. of dehydrated ethanol.After the precipitated NaCl is filter otf, 1.46 g. of diethyl oxalate isadded into the filtrate and allowed to stand overnight. The resultingprecipitate is collected by suction. Yield 1.13 g. (62%), M.P. 266267 C.(with decomposition). Analysis.Calcd. for C H O N C, 39.34; H, 4.95; N,38.28. Found: C, 39.61; H, 4.90; N, 38.32.

The following examples are illustrative of the preparation of variouscompounds represented by Formula I, above:

EXAMPLE 9 Preparation of4-an1in0-6-is0pr0pylamina-strz'azine-Z-carban.'lide c rr on N, 30.80.Found: N, 30.80.

7 EXAMPLE 10 Preparation of 4-amino-6-cyclohexylamino-s-triazine-Z-carboxypiperidide A mixture of 2.37 g. of 1-cyclohexyl-3-(4,5-dioxo-2-imidazolidinylidene)-guanidine and 1 cc. of piperidine is heated for 1hour on a water bath. The resulting precipitate is collected by suctionon cooling and recrystallized from ethanol to colorless prisms whichmelt at 266 C.; yield 1.64 g. (54%). Analysis.Calcd. for C H ON N,27.61. Found: N, 27.89.

EXAMPLE 11 Preparation of 4-amino-6-(p-met/zoxyphenylamino)-s-triazine-Z-carboxypyrrolidide A mixture of 2.61 g. of1-(p-methoxyphenyl)-3-(4,5- dioxo-2-imi'dazolidinylidene)-guanidine and1 cc. of pyrrolidine is refluxed for 1 hour on a water bath. Aftercooling, the deposited precipitate is collected by filtration andrecrystallized from ethanol to colorless needles which melt at 215-216C.; yield 1.82 g. (58%). Analysis.- Calcd. for C H O N N, 26.74. Found:N, 26.73.

EXAMPLE 12 Preparation of 4-amino-6-(N-morpholinyl)-s-triazine2-carb0xyguanidide hydrate Preparation of4-amin0-6-(p-toluidino)-s-triazine- Z-carboxypyrrolidide A mixture of2.45 g. of 1-(p-tolyl)-3-(4,5-dioxo-2- imidazolidinylidene)-guanidineand 0.71 g. of pyrrolidine is heated for 2 hours on a Water bath. Aftera completion of the reaction, the resulting residue is recrystallizedfrom a small amount of ethanol to give colorless prisms which melt at258 C.; yield 1.24 g. (50%). Analysis. Calcd. for C H ON N, 28.17.Found: N, 28.34.

EXAMPLE 14 Preparation of 4-amz'n0-6-(p-toluidino)-s-triazine2-carb0xylic acid Procedure 1.2.45 g. of 1-(p-tolyl)-3-(4,5-dioxo2-imidazolidinylidene)-guanidine is dissolved in a dilute alkaline aqueoussolution and the solution is neutralized with hydrochloric acid. Theresulting precipitate is collected by suction, washed with hot water anddried at C. under pressure; MP. 234235 C. (with decomposition).Analysis.Calcd. for C H O N C, 53.87; H, 4.52; N, 28.56. Found: C,53.21; H, 4.29; N, 28.48.

Procedure 2.2.7 g. of ethyl 4-arnino-6-(p-toluidino)-s-triazine-2-carboxylate is refluxed with ethanolic sodium hydroxidesolution for 5 hours. After cooling, the precipitate is collected anddissolved in a small amount of Water, followed by neutralization withhydrochloric acid, and the colorless crystals precipitated are collectedby suction, M.P. 234235 C. 7

Procedure 3.A mixture of 1.91 g. of 1-(p-tolyl)- biguanide and 1.51 g.of potassium salt of monethyl oxalate in dehydrated ethanol is refluxedfor 10 hours. When cool, the precipitate is filtered and dissolved in asmall amount of water. The solution is neutralized with hydrochloricacid and the colorless crystals precipitated are collected by suction,MP. 234-235 C.

EXAMPLE 15 Procedure 1.Preparati0n of ethyl-4-amino-6 (N -m0rph0linyl-s-triazine-2-carb0xy late 2.25 g. of1,1-(2,2-oxydiethyl)-3-(4,5-dioxo-2-imidazolidinylidene)-guanidine incc. of dehydrated ethanol is refluxed for 10 hours. The mixture isconcentrated, followed by chilling. The precipitate is filtered andrecrystallized from ethanol to obtain colorless prisms; yield 1.62 g.,MP. 172 C. Analysis.Calcd. for C H O N N, 27.66. Found: N, 28.13.

Procedure 2.-Preparati0n of4-amino-6-(N-morpholinyl)-s-triazine-2-carb0xyguanidide To a solution of2.53 g. of ethyl 4-amino-6-(N-mor pholinyl)-s-triazine-2-carboxylate in100 cc. of dehydrated ethanol, 0.6 g. of guanidine Was added. Afterrefluxing on a water bath for 5 hours, the mixture was concentrated andchilled. The precipitate was collected by suction, recrystallized fromwater to colorless prisms; M.P. 271 C.

EXAMPLE 16 Preparation of 4-amino-6-(p-t0luidino)-s-triazine-Z-cm'b0xypyrr0lidide To 2.45 g. of4-amino-6-(p-toluidino)-s-triazine-2- carboxyl-ic acid prepared by theabove method is added an excess amount of thionyl chloride. Afterremoval of spectively, there are obtained, instead of2-amino-4-morpholino-6-(N-amidinocarbamoyl)-s-triazine as the finalproduct, the corresponding 2,4-diamino-6- (Naamidinocarb amoyl)-s-triazine,

2-amino-4-N-methy1amino-6- (N-amidinocarbamoyl) -striazine,

Z-amino-4-N-ethy1amino-6- (N-amidinocarbamoyl) -stri-azine,

2-amino-4- (N-methyl-N-isopropylamino) -6- (N-amidinocarb amoyl-s-triazine,

2-amino-4-N-n-butylamino-6- (N-amidino carbamoyl -sttriaz-ine,

2-amino-4-isobutylamino-6- (N-amidinocarbamoyl) -striazine,

2-amino-4- (N-pentyl-N-t-butyl-amino) -6-(N-amidinocarbamoy1)-s-triazine,

2-arnino-4-heXylamino-6- (N-amidinocarbamoyl) -striazine,

2-amino-4- 2-ethy1hexy1amino -6- (N-amidinocarbamoyl -s-triazine,

2-amino-4-piperidino-6- (N-amidinocarb amoyl -striazine,

2-amino-4-pyrro'1idino-6- (N-amidinocarbamoyl) -striazine, and

2-amino-4-cyclohexylamino-6- (N-amidino carbamoyl -striazine.

12 3. A compound selected from the group consisting of compounds of theformula wherein the group R1 and R taken together with N- constitute aheterocyclic group selected from the group consisting of morpholino,piperidino and pyrrolidino, and X is a member selected from the groupconsisting of chloro, bromo, fluor, iodo, and (lower)alkoxy.

References Cited by the Examiner UNITED STATES PATENTS 4/1959 Bellet260247.2 10/1962 Strassberger et a1. 260-2499 OTHER REFERENCESSokolovskara et 211.: Chemical Abstracts, vol. 51, page 16, 493 (1957).

ALEX MAZEL, Primary Examiner.

NICHOLAS RIZZO, HENRY R. JILES, JOSE TOVAR,

Assistant Examiners.

3. A COMPOUND SELECTED FROM THE GROUP CONSISTING OF COMPOUNDS OF THEFORMULA